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    Amonafide (AS1413)
    Amonafide (AS1413)

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V1417
    CAS #: 69408-81-7Purity ≥98%

    Description: Amonafide (formerly NSC308847, AS1413; AS-1413; NSC-308847; Amonafide; Nafidimide; Benzisoquinolinedione; Quinamed; Xanafide), an imide derivative of naphthalic acid, is an inhibitor of topoisomerase II (Topo II) and also a DNA intercalator being investigated for the treatment of cancer.  

    References: Cancer Res. 1987 Feb 15;47(4):1040-4; Expert Rev Hematol. 2012 Feb;5(1):17-26.

    Related CAS #: 69408-81-7 (free base)   150091-68-2 (2HCl)   618863-54-0 (L-malate)   618863-60-8 

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    Molecular Weight (MW)283.33 
    CAS No.69408-81-7 
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 57 mg/mL (201.2 mM) 
    Water: <1 mg/mL
    Ethanol: 4 mg/mL (14.1 mM) 
    Other infoChemical Name: 5-Amino-2-[2-(dimethylamino)ethyl]benzo[de]isoquinoline-1,3-dione
    InChi Code: InChI=1S/C16H17N3O2/c1-18(2)6-7-19-15(20)12-5-3-4-10-8-11(17)9-13(14(10)12)16(19)21/h3-5,8-9H,6-7,17H2,1-2H3
    SMILES Code: O=C1N(CCN(C)C)C(C2=CC(N)=CC3=CC=CC1=C23)=O
    SynonymsAS1413; NSC308847, AS1413; NSC-308847, AS-1413; NSC 308847, AS 1413; Amonafide; Nafidimide; Benzisoquinolinedione; Quinamed; Xanafide

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    In Vitro

    In vitro activity: Through a topoisomerase II-mediated reaction, Amonafide treatment produces DNA single-strand breaks (SSB), double-strand breaks (DSB), and DNA-protein cross-links in human myeloid leukemia cells. Amonafide treatment inhibits conlony formation of the leukemic cell lines and the normal human bone marrow GM-CFC in a dose-dependent manner. Amonafide does not produce topoisomerase I-mediated DNA cleavage even at 100 μM. The m-AMSA-resistant line is less than 2-fold resistant to Amonafide. Amonafide interferes with the DNA breakage-reunion activity of mammalian DNA topoisomerase II resulting in DNA cleavage stimulation. Compared with those of other antitumor drugs, Amonafide-stimulated cleavage intensity patterns are markedly different. Amonafide highly prefers a cytosine, and excludes guanines and thymines instead, at position -1, with lower preference for an adenine at position +1. Topoisomerase II-mediated DNA cleavage induced by Amonafide is affected only slightly (less than 3-fold) by 1 mM ATP, suggeting that Amonafide is an ATP-insensitive topoisomerase II inhibitor in contrast to doxorubicin, etoposide, and mitoxantrone. Amonafide significantly inhibits the growth of HT-29, HeLa, and PC3 cells with IC50 of 4.67 μM, 2.73 μM, and 6.38 μM, respectively. Amonafide is unaffected by P-glycoprotein-mediated efflux, unlike those of the classical topoisomerase II inhibitors (daunorubicin, doxorubicin, idarubicin, etoposide, and mitoxantrone).

    Cell Assay: All cell lines (HT-29, HeLa, and PC3) are in the logarithmic phase of growth when the assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) is carried out. Cells are harvested and seeded into 96-well tissue culture plates at a density of 2.5 × 103 cells/well in 150 μL aliquots of medium. The concentrations tested are serial dilutions of a stock solution (10 μM in DMSO) with phosphate-buffered saline (PBS) and are added 24 hours later. The assay is ended after 72 hours of Amonafide exposure and PBS is used as a negative control. After 72 hours treatment, cells are washed twice with PBS, and then 50 μL/well of MTT reagent (1 mg/mL in PBS) together with 150 μL/well of prewarmed medium are added. The plates are returned to the incubator for 4 hours. Subsequently, DMSO is added as solvent. Absorbance is determined at 570 nm with a Microplate reader. All experiments are performed at least three times, and the average of the percentage absorbance is plotted against concentration. Then, the concentration of Amonafide required to inhibit 50% of cell growth (IC50) is calculated for Amonafide.

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    ReferencesCancer Res. 1987 Feb 15;47(4):1040-4; Expert Rev Hematol. 2012 Feb;5(1):17-26. 

    These protocols are for reference only. InvivoChem does not independently validate these methods.


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